Use of DEAE-cellulose in the separation of proteins from egg white and other biological materials

1960 ◽  
Vol 3 ◽  
pp. 256-264 ◽  
Author(s):  
Stanley Mandeles
Keyword(s):  
Deae Cellulose ◽  
Biological Materials ◽  
Egg White ◽  
Separation Of Proteins

OVALBUMIN AND OTHER WATER-SOLUBLE PROTEINS IN AVIAN YOLK DURING EMBRYOGENESIS

1966 ◽  
Vol 44 (3) ◽  
pp. 293-301 ◽  
Author(s):  
Z. Saito ◽  
W. G. Martin
Keyword(s):  
Phosphorus Content ◽  
Great Increase ◽  
Authentic Sample ◽  
Deae Cellulose ◽  
Egg Yolk ◽  
Egg White ◽  
Water Soluble ◽  
The Other ◽  
Water Soluble Protein ◽  
Soluble Material

The proportion of water-soluble protein in egg yolk increased markedly after 12 days' incubation, concurrent with the appearance of a new fraction that evidently entered from the egg white. The electrophoretic, sedimentation [Formula: see text], molecular weight (45 × 103), and chromatographic properties of this new fraction, after 18 days' incubation, were the same as, and the tyrosine–tryptophane and phosphorus–nitrogen ratios were similar to those of authentic ovalbumin. Ovalbumin is known to have two components differing in phosphorus content and both were evident in the authentic sample and in the new yolk fraction. At the later stages of incubation the ovalbumin component of lowest phosphorus content predominated and this also may be the form that was present in the serum of the embryo. Changes in the other components of the water-soluble material during embryogenesis were less dramatic. Chromatography on DEAE-cellulose revealed at least nine yolk components in unincubated egg, although only the three livetins were made evident by free-boundary electrophoresis. The great increase in ovalbumin prevented detection of changes in the livetins during embryogenesis but α-livetin (serum albumin) apparently disappeared most rapidly and completely.

New Design for a Differentially Pumped Hydration Chamber for a Siemens IA

1971 ◽  
Vol 29 ◽  
pp. 44-45
Author(s):  
R. C. Moretz ◽  
G. G. Hausner ◽  
D. F. Parsons
Keyword(s):  
Electron Microscopy ◽  
Water Vapor ◽  
Electron Diffraction ◽  
Water Vapor Pressure ◽  
Biological Materials ◽  
Thin Membrane ◽  
Reliable System ◽  
System A ◽  
Water Films ◽  
Aperture Opening

Electron microscopy and diffraction of biological materials in the hydrated state requires the construction of a chamber in which the water vapor pressure can be maintained at saturation for a given specimen temperature, while minimally affecting the normal vacuum of the remainder of the microscope column. Initial studies with chambers closed by thin membrane windows showed that at the film thicknesses required for electron diffraction at 100 KV the window failure rate was too high to give a reliable system. A single stage, differentially pumped specimen hydration chamber was constructed, consisting of two apertures (70-100μ), which eliminated the necessity of thin membrane windows. This system was used to obtain electron diffraction and electron microscopy of water droplets and thin water films. However, a period of dehydration occurred during initial pumping of the microscope column. Although rehydration occurred within five minutes, biological materials were irreversibly damaged. Another limitation of this system was that the specimen grid was clamped between the apertures, thus limiting the yield of view to the aperture opening.

Evaluation of Freezing Methods by Low Temperature X-Ray Diffraction

1977 ◽  
Vol 35 ◽  
pp. 330-333
Author(s):  
T. Gulik-Krzywicki ◽  
M.J. Costello
Keyword(s):  
Biological Materials ◽  
Molecular Organization ◽  
X Ray Diffraction ◽  
Glass Capillary ◽  
X Ray ◽  
Rate Dependent ◽  
Before And After ◽  
Freeze Etching ◽  
Diffraction Patterns ◽  
Set Up

Freeze-etching electron microscopy is currently one of the best methods for studying molecular organization of biological materials. Its application, however, is still limited by our imprecise knowledge about the perturbations of the original organization which may occur during quenching and fracturing of the samples and during the replication of fractured surfaces. Although it is well known that the preservation of the molecular organization of biological materials is critically dependent on the rate of freezing of the samples, little information is presently available concerning the nature and the extent of freezing-rate dependent perturbations of the original organizations. In order to obtain this information, we have developed a method based on the comparison of x-ray diffraction patterns of samples before and after freezing, prior to fracturing and replication.Our experimental set-up is shown in Fig. 1. The sample to be quenched is placed on its holder which is then mounted on a small metal holder (O) fixed on a glass capillary (p), whose position is controlled by a micromanipulator.

Enhanced information from biological materials through technological improvements in cryo-electron microscopy

1992 ◽  
Vol 50 (1) ◽  
pp. 788-789
Author(s):  
Marc J.C. de Jong ◽  
Wim M. Busing ◽  
Max T. Otten
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
Biological Materials ◽  
Electron Crystallography ◽  
Cryo Electron Microscopy ◽  
Transmission Electron ◽  
The Many ◽  
Technological Improvements ◽  
Beam Damage

Biological materials damage rapidly in the electron beam, limiting the amount of information that can be obtained in the transmission electron microscope. The discovery that observation at cryo temperatures strongly reduces beam damage (in addition to making it unnecessaiy to use chemical fixatives, dehydration agents and stains, which introduce artefacts) has given an important step forward to preserving the ‘live’ situation and makes it possible to study the relation between function, chemical composition and morphology.Among the many cryo-applications, the most challenging is perhaps the determination of the atomic structure. Henderson and co-workers were able to determine the structure of the purple membrane by electron crystallography, providing an understanding of the membrane's working as a proton pump. As far as understood at present, the main stumbling block in achieving high resolution appears to be a random movement of atoms or molecules in the specimen within a fraction of a second after exposure to the electron beam, which destroys the highest-resolution detail sought.

Application of ruthenium red/osmium tetroxide to bacteria, plant and animal tissue

1986 ◽  
Vol 44 ◽  
pp. 54-55
Author(s):  
R. L. Grayson ◽  
N. A. Rechcigl
Keyword(s):  
Electron Microscopy ◽  
Electron Density ◽  
Osmium Tetroxide ◽  
Biological Materials ◽  
Ruthenium Red ◽  
Animal Tissue

Ruthenium red (RR), an inorganic dye was found to be useful in electron microscopy where it can combine with osmium tetroxide (OsO4) to form a complex with attraction toward anionic substances. Although Martinez-Palomo et al. (1969) were one of the first investigators to use RR together with OsO4, our computor search has shown few applications of this combination in the intervening years. The purpose of this paper is to report the results of our investigations utilizing the RR/OsO4 combination to add electron density to various biological materials. The possible mechanisms by which this may come about has been well reviewed by previous investigators (1,3a,3b,4).

Three-Dimensional Structure of Cloned T3 Connector Protein at 1.6nm Resolution

1990 ◽  
Vol 48 (1) ◽  
pp. 282-283
Author(s):  
José L. Carrascosa ◽  
José M. Valpuesta ◽  
Hisao Fujisawa
Keyword(s):  
Dna Sequences ◽  
Expression Vector ◽  
Three Dimensional ◽  
Deae Cellulose ◽  
Dna Packaging ◽  
Dimensional Structure ◽  
Two Dimensional ◽  
Freezing And Thawing ◽  
Three Dimensional Structure ◽  
Dimensional Reconstruction

The head to tail connector of bacteriophages plays a fundamental role in the assembly of viral heads and DNA packaging. In spite of the absence of sequence homology, the structure of connectors from different viruses (T4, Ø29, T3, P22, etc) share common morphological features, that are most clearly revealed in their three-dimensional structure. We have studied the three-dimensional reconstruction of the connector protein from phage T3 (gp 8) from tilted view of two dimensional crystals obtained from this protein after cloning and purification.DNA sequences including gene 8 from phage T3 were cloned, into Bam Hl-Eco Rl sites down stream of lambda promotor PL, in the expression vector pNT45 under the control of cI857. E R204 (pNT89) cells were incubated at 42°C for 2h, harvested and resuspended in 20 mM Tris HC1 (pH 7.4), 7mM 2 mercaptoethanol, ImM EDTA. The cells were lysed by freezing and thawing in the presence of lysozyme (lmg/ml) and ligthly sonicated. The low speed supernatant was precipitated by ammonium sulfate (60% saturated) and dissolved in the original buffer to be subjected to gel nitration through Sepharose 6B, followed by phosphocellulose colum (Pll) and DEAE cellulose colum (DE52). Purified gp8 appeared at 0.3M NaCl and formed crystals when its concentration increased above 1.5 mg/ml.

Biological Materials Science

2014 ◽  
Author(s):  
Marc Andre Meyers ◽  
Po-Yun Chen
Keyword(s):  
Materials Science ◽  
Biological Materials
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